The method is non-invasive laboratory confirmation of immunoglobulin-e (ige)-dependent genesis seasonal rhinoconjunctivitis (ibs) in children

 

(57) Abstract:

The invention relates to medicine, in particular to the field of Allergology and can be used for confirmation of IgE-dependent Genesis of IBS in children. The inventive method is carried out as follows: the child take the saliva, by ELISA to determine the levels of total IgE using a set of reagents for the quantitative determination of total IgE in serum, plasma and amniotic fluid, and when the values of total IgE in saliva than 1.0 IU/ml diagnose the presence of IgE-dependent Genesis of IBS in children. The advantage of the invention lies in the fact that the use as material for the study of saliva allows painless for the child to be examined without damaging his psyche. 3 table.

The invention relates to medicine, in particular to the field of Allergology and can be used to confirm atopic Genesis of IBS in children. There is a method of confirmation of IgE-dependent Genesis of IBS in children in the serum (Jaeger, L. (ed) Clinical immunology and Allergology in 3 volumes. - M.: Medicine, 1990, I. 2. - S. 50-51, 58-59; Hamilton R. G., N. Franklin Adkinson Jr. Assessment of human allergic diseases. In book Clinical immunology: principals and practice. - ed. - in chief R. Rich, ed. T. A. Fleisher, B. D. Schwartz, W. T. Shearer, W. Strober. omicheskogo disease, although normal levels of total IgE does not exclude it. Approximately 50% of patients with allergic rhinitis with seasonal exacerbations is determined normal level of serum total IgE (Report of an international consensus on the diagnosis and treatment of rhinitis // Russian rhinology. - 1996. No. 4. - S. 21). Determination of total IgE in serum is shown when carrying out differential diagnosis of allergic rhinitis with his other forms (Lawlor, (as amended) Clinical immunology and Allergology. - M.: “The Practice,” 2000. - S. 54). There are about 18 methods and their modifications for the determination of total IgE in serum (Hamilton R. G., N. Franklin Adkinson Jr. Assessment of human allergic diseases. In book Clinical immunology: principals and practice. - ed. - in chief R. Rich, ed. T. A. Fleisher, B. D. Schwartz, W. T. Shearer, W. Strober. - 1996. - vol. 2. - St. Louis, Mosby. -R. 2176). When this occurs invasive nature of the sampling of the material for research.

There is a method of confirmation of IgE - dependent Genesis of IBS in children in the serum, in particular, the enzyme-linked immunosorbent assay (ELISA) (Hamilton R. G., N. Franklin Adkinson, Jr. Assessment of human allergic diseases. In book Clinical immunology: principals and practice. - ed. - in chief R. Rich, ed. T. A. Fleisher, B. D. Schwartz, W. T. Shearer, W. Strober. - 1996. - vol. 2. - St. Louis, Mosby. - P. 2176), selected as a prototype.

The known method is that take blood from a vein, CSOs IgE, in excess of normal levels, confirm IgE-dependent Genesis of IBS in children. The disadvantages of this method are that for confirmation of the diagnosis it is necessary to take blood from a vein that causes the pain, causing the child fear survey is causing a kind of psychological trauma for the patient, requires certain technical skills of medical staff is a potential risk of infection, it has disadvantages invasive nature of the sampling of the material. In addition, the level of total IgE serum gives an estimate of the systemic immune system, while the disease pathogenesis of IBS is associated mainly with local changes of IgE production at the level of the mucous membranes of the upper respiratory tract, and therefore the level of IgE in the saliva may have greater diagnostic value.

The goal towards which the inventive method is directed, is the elimination of the disadvantages invasive nature of the sampling of the material, i.e. removal of the patient's fear, pain, risk of infection, as well as improving the accuracy of diagnosis of local accumulation of IgE level mucosaassociated lymphoid tissue, due to the transition to non-invasive way to confirm IgE-sovietinvasion confirmation of IgE-dependent Genesis of IBS in children, the patient takes the material to study and determine the level of total IgE by ELISA using standard commercial test systems for the determination of IgE in serum (plasma) blood and amniotic fluid and at values higher than normal conclude about IgE-dependent Genesis of IBS in children, thus according to the invention as a material for research use saliva.

Use as material for the study of saliva allows painless for the child to be examined without damaging his psyche. There is no limitation of the scope of the investigated material is easier to obtain the necessary volume. The results of the research allow saliva to evaluate the local accumulation of IgE.

The inventive method has the novelty in comparison with the prototype, it is the fact that as the material for research use saliva, which allows to estimate the local accumulation of IgE using the same test system, which is used in the study of blood.

The applicant is not known to the confirmation of IgE-dependent Genesis of IBS in children by saliva, so he believes that the claimed technical solution meets the criterion of “inventive step”.

The inventive method non-invasive laboratory confirmation of IgE-dependent Genesis of IBS in children can be widely used in medicine, namely in Allergology and therefore meets the criterion of “industrial application”.

The inventive method is carried out as abrup" reagents for the quantitative determination of total IgE in serum, plasma and amniotic fluid (JSC "Vector - BEST, Novosibirsk region, p. Koltsovo) according to the instructions of the reagent kit.

Principle of the ELISA method described (Hamilton R. G., N. Franklin Adkinson, Jr. Assessment of human allergic diseases. In book Clinical immunology: principals and practice. - ed. - in chief R. Rich, ed. T. A. Fleisher, B. D. Schwartz, W. T. Shearer, W. Strober. - 1996. - vol. 2. - St. Louis, Mosby. - P. 2176). The study of saliva is carried out in accordance with the instructions for the kit "IgE - ELISA - BEST - strip" for the quantitative determination of total IgE in serum, plasma and amniotic fluid JSC "Vector - BEST, Novosibirsk region, p. Koltsov. Used solid-phase immunoassay method based on the "sandwich" principle. The analysis is carried out in two stages. In the first stage of the calibration sample with a known concentration of IgE and analyzed samples are incubated in the wells stripyoung tablet with immobilized monoclonal antibodies (MCAT) to IgE. The tablet then washed. In the second stage of bound peroxidase in the hole IgE treated with a conjugate of MCAT to IgE with horseradish peroxidase (conjugate of MCAT and immobilized in the wells of MCAT specific to different parts of the molecule IgE). After washing off the excess conjugate is formed immune complexes immobilized MCAT - IgE conjugate reveal tensively coloring Chromogen is proportional to the concentration of IgE in the analyzed sample. After stopping the reaction with stop reagent results are recorded photometrically. Measurement of total IgE in IU/ml carried out on a tablet Multiscan photometer plus (Labsystems), the filter of 492 nm. Values of total IgE in saliva than 1.0 IU/ml testify in favor of the existence of IgE-dependent Genesis of IBS in children. Examined children 7-16 years. The authors found that in the saliva of healthy children, the level of total IgE does not exceed 1.0 IU/ml, and in the saliva of patients with IBS children the level of total IgE significantly increased in the period of remission and exacerbation of the disease (table 1) compared with healthy children of this age, diagnostic significant level values are greater than 1.0 IU/ml

Indicators of the significance of diagnostic confirmation method IgE-dependent Genesis of IBS in children are presented in table 2, where the values of sensitivity, specificity, efficiency, diagnostic test, and predictive value of positive and negative tests. The method allows to confirm IgE-dependent development path IBS is a non-invasive method in the period of remission and exacerbation. The method is not inferior in its sensitivity and specificity generally accepted method of determining the level of total IgE in serum in ps.ru/fullimg4/rupat5/2004/09/03/2237250-2t.gif" ALIGN="ABSMIDDLE">

In comparison with the prototype of the proposed method has the advantages of non-invasive method that is painless, does not cause fear of examination, the child has no potential risk of infection, has simplicity in obtaining the necessary material, a sufficient amount of material for research, allows to estimate the local accumulation of IgE level mucosaassociated lymphoid tissue.

The method is non-invasive laboratory confirmation immunoglobulin E(YgE) - dependent Genesis seasonal Rhino conjunctivitis (IBS) in children, namely, that the patient take the material to study and determine the level of General YgE enzyme-linked immunosorbent assay (ELISA) using standard commercial test systems for determining YgE, characterized in that as the material for research use saliva and at values above 1.0 IU/ml conclude about YgE - dependent Genesis of IBS in children.

 

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